{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/139821"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/139821","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Molecular characterization of soybean meal trypsin inhibitors and lectins as a basis for developing approaches to mitigate their anti-nutritional effects","abstract":"Soybean is a crop that is widely eaten because of its nutritive properties. However, soybean cannot be consumed in its raw form because it contains anti-nutrients like trypsin inhibitors which bind to serine proteases (chymotrypsin, elastase and trypsin) and restrain their digestive functions leading to indigestion and stunted growth. It also contains lectin which binds to intestinal cells containing N-acetyl galactosamine (GalNAc) to impede nutrient uptake into the blood. Soybean needs to be processed to get rid of these anti-nutrients before human or animal consumption. The conventional method of soybean processing which involves the moist heat treatment of soybeans is time and energy demanding, leads to the loss of nutrients, and residual amount of anti-nutrients are retained in the processed soybean meal, making this process relatively inefficient. This research work aimed to provide alternative solutions to the conventional soybean processing methods. Using quick purification techniques, we isolated TIs [Bowman-Birk trypsin inhibitor (BBTI) and Kunitz trypsin inhibitor (KTI)] and lectin from soybean meal. Using biophysical techniques, we characterized the interactions between soybean anti-nutrients and their host ligands to offer alternative solutions to improve the nutritional quality of soybean meal. The known anti-nutritional pathway of soybean lectin involves its binding to GalNAc containing intestinal cells; however, we believe soybean lectin could be also targeting another ligand called sulfatide besides GalNAc in the small intestine due to some similarities between soybean lectin and galectin-4, a mammalian lectin that binds to sulfatide in the small intestine. Hence, we also explored soybean lectin to sulfatide interactions to find a new soybean lectin anti-nutritional pathway. Results from a soybean meal cultivar showed that KTI had a binding preference for chymotrypsin, while BBTI preferred binding to trypsin and elastase. This provides insights into the unique roles that both TIs play in soybean and why the plant retains both. After screening TIs from several soybean meal lines for their affinity to trypsin, we identified BBTI soybean lines with ~4 to 6-fold lower affinity for trypsin and a KTI soybean line with ~8-fold lower affinity for trypsin, compared to the standard BBTI and KTI, respectively. These promising soybean meal lines have been designed for soybean crossbreeding to produce hybrid soybean meal lines that would require little to no processing before animal consumption. We also identified the KTI amino acid sequence motif, SPLHALFI as a suitable motif for gene editing to produce soybean meal lines with improved nutritional qualities. After screening GalNAc and GalNAc analogs against lectin to find a higher lectin affinity ligand than GalNAc that could be introduced as an additive in soybean meal to bind to lectin and prevent lectin from binding to GalNAc containing intestinal cells, we found no higher affinity ligand than GalNAc for lectin. However, we found out that a suitable substitution on the anomeric carbon of GalNAc would lead to the design of higher lectin affinity ligands than GalNAc that could serve as additives in soybean meal to mitigate lectin adverse effects. We also found out that soybean lectin crosslinks and binds to sulfatides with a high affinity, in a process that could result in unwanted signaling events in the body, making this another soybean lectin antinutritional pathway. This discovery could also lead to the design of suitable additives to target the sulfatide binding site on soybean lectin and prevent soybean lectin from binding to sulfatide in the body. It is recommended that this newly discovered soybean lectin-sulfatide anti-nutritional pathway should also be considered when assessing the nutritional quality of soybean meal. Taken together, this research provides alternative solutions that would lead to the enhancement of the nutritional quality of soybean meal and the mitigation of the adverse effects of soybean's anti-nutrients.","abstract_html":"Soybean is a crop that is widely eaten because of its nutritive properties. However, soybean cannot be consumed in its raw form because it contains anti-nutrients like trypsin inhibitors which bind to serine proteases (chymotrypsin, elastase and trypsin) and restrain their digestive functions leading to indigestion and stunted growth. It also contains lectin which binds to intestinal cells containing N-acetyl galactosamine (GalNAc) to impede nutrient uptake into the blood. Soybean needs to be processed to get rid of these anti-nutrients before human or animal consumption. The conventional method of soybean processing which involves the moist heat treatment of soybeans is time and energy demanding, leads to the loss of nutrients, and residual amount of anti-nutrients are retained in the processed soybean meal, making this process relatively inefficient. This research work aimed to provide alternative solutions to the conventional soybean processing methods. Using quick purification techniques, we isolated TIs [Bowman-Birk trypsin inhibitor (BBTI) and Kunitz trypsin inhibitor (KTI)] and lectin from soybean meal. Using biophysical techniques, we characterized the interactions between soybean anti-nutrients and their host ligands to offer alternative solutions to improve the nutritional quality of soybean meal. The known anti-nutritional pathway of soybean lectin involves its binding to GalNAc containing intestinal cells; however, we believe soybean lectin could be also targeting another ligand called sulfatide besides GalNAc in the small intestine due to some similarities between soybean lectin and galectin-4, a mammalian lectin that binds to sulfatide in the small intestine. Hence, we also explored soybean lectin to sulfatide interactions to find a new soybean lectin anti-nutritional pathway. Results from a soybean meal cultivar showed that KTI had a binding preference for chymotrypsin, while BBTI preferred binding to trypsin and elastase. This provides insights into the unique roles that both TIs play in soybean and why the plant retains both. After screening TIs from several soybean meal lines for their affinity to trypsin, we identified BBTI soybean lines with ~4 to 6-fold lower affinity for trypsin and a KTI soybean line with ~8-fold lower affinity for trypsin, compared to the standard BBTI and KTI, respectively. These promising soybean meal lines have been designed for soybean crossbreeding to produce hybrid soybean meal lines that would require little to no processing before animal consumption. We also identified the KTI amino acid sequence motif, SPLHALFI as a suitable motif for gene editing to produce soybean meal lines with improved nutritional qualities. After screening GalNAc and GalNAc analogs against lectin to find a higher lectin affinity ligand than GalNAc that could be introduced as an additive in soybean meal to bind to lectin and prevent lectin from binding to GalNAc containing intestinal cells, we found no higher affinity ligand than GalNAc for lectin. However, we found out that a suitable substitution on the anomeric carbon of GalNAc would lead to the design of higher lectin affinity ligands than GalNAc that could serve as additives in soybean meal to mitigate lectin adverse effects. We also found out that soybean lectin crosslinks and binds to sulfatides with a high affinity, in a process that could result in unwanted signaling events in the body, making this another soybean lectin antinutritional pathway. This discovery could also lead to the design of suitable additives to target the sulfatide binding site on soybean lectin and prevent soybean lectin from binding to sulfatide in the body. It is recommended that this newly discovered soybean lectin-sulfatide anti-nutritional pathway should also be considered when assessing the nutritional quality of soybean meal. Taken together, this research provides alternative solutions that would lead to the enhancement of the nutritional quality of soybean meal and the mitigation of the adverse effects of soybean's anti-nutrients.","abstract_has_math":false,"creators":["Okedigba, Ayoyinka Oluwaseun"],"institution":"Virginia Tech","degree_name":"Doctor of Philosophy","degree_level":"doctoral","degree_discipline":"Chemistry","degree_department":"Chemistry","school":null,"contributors":[],"advisors":[],"committee_chairs":["Capelluto, Daniel G."],"committee_members":["Schubot, Florian David","Welborn, Valerie","Mevers, Emily Elizabeth"],"year":2025,"date_issued":"2025-12-03","date_published":"2025-12-03","updated_at":"2026-07-22T22:19:42Z","subjects":["Soybean meal","Bowman-Birk trypsin inhibitor","Kunitz trypsin inhibitor","serine proteases","lectin","N-acetyl galactosamine","sulfatides"],"languages":["en"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["vt_gsexam:44800"],"render_values":[{"text":"vt_gsexam:44800","href":null,"code":true}]}]},"links":{"outbound_url":"https://hdl.handle.net/10919/139821","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.committeechair","label":"Committee Chair","values":["Capelluto, Daniel G."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Schubot, Florian David","Welborn, Valerie","Mevers, Emily Elizabeth"]},{"key":"dc:contributor.department","label":"Department","values":["Chemistry"]},{"key":"dc:creator","label":"Author","values":["Okedigba, Ayoyinka Oluwaseun"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2025-12-04T09:00:22Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2025-12-04T09:00:22Z"]},{"key":"dc:date.issued","label":"Date","values":["2025-12-03"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Tech"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["doctoral"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Virginia Polytechnic Institute and State University"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Soybean meal","Bowman-Birk trypsin inhibitor","Kunitz trypsin inhibitor","serine proteases","lectin","N-acetyl galactosamine","sulfatides"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["In Copyright"]},{"key":"dc:rights.uri","label":"Rights URI","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.other","label":"Dc Identifier Other","values":["vt_gsexam:44800"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/10919/139821"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Soybean is a crop that is widely eaten because of its nutritive properties. However, soybean cannot be consumed in its raw form because it contains anti-nutrients like trypsin inhibitors which bind to serine proteases (chymotrypsin, elastase and trypsin) and restrain their digestive functions leading to indigestion and stunted growth. It also contains lectin which binds to intestinal cells containing N-acetyl galactosamine (GalNAc) to impede nutrient uptake into the blood. Soybean needs to be processed to get rid of these anti-nutrients before human or animal consumption. The conventional method of soybean processing which involves the moist heat treatment of soybeans is time and energy demanding, leads to the loss of nutrients, and residual amount of anti-nutrients are retained in the processed soybean meal, making this process relatively inefficient. This research work aimed to provide alternative solutions to the conventional soybean processing methods. Using quick purification techniques, we isolated TIs [Bowman-Birk trypsin inhibitor (BBTI) and Kunitz trypsin inhibitor (KTI)] and lectin from soybean meal. Using biophysical techniques, we characterized the interactions between soybean anti-nutrients and their host ligands to offer alternative solutions to improve the nutritional quality of soybean meal. The known anti-nutritional pathway of soybean lectin involves its binding to GalNAc containing intestinal cells; however, we believe soybean lectin could be also targeting another ligand called sulfatide besides GalNAc in the small intestine due to some similarities between soybean lectin and galectin-4, a mammalian lectin that binds to sulfatide in the small intestine. Hence, we also explored soybean lectin to sulfatide interactions to find a new soybean lectin anti-nutritional pathway. Results from a soybean meal cultivar showed that KTI had a binding preference for chymotrypsin, while BBTI preferred binding to trypsin and elastase. This provides insights into the unique roles that both TIs play in soybean and why the plant retains both. After screening TIs from several soybean meal lines for their affinity to trypsin, we identified BBTI soybean lines with ~4 to 6-fold lower affinity for trypsin and a KTI soybean line with ~8-fold lower affinity for trypsin, compared to the standard BBTI and KTI, respectively. These promising soybean meal lines have been designed for soybean crossbreeding to produce hybrid soybean meal lines that would require little to no processing before animal consumption. We also identified the KTI amino acid sequence motif, SPLHALFI as a suitable motif for gene editing to produce soybean meal lines with improved nutritional qualities. After screening GalNAc and GalNAc analogs against lectin to find a higher lectin affinity ligand than GalNAc that could be introduced as an additive in soybean meal to bind to lectin and prevent lectin from binding to GalNAc containing intestinal cells, we found no higher affinity ligand than GalNAc for lectin. However, we found out that a suitable substitution on the anomeric carbon of GalNAc would lead to the design of higher lectin affinity ligands than GalNAc that could serve as additives in soybean meal to mitigate lectin adverse effects. We also found out that soybean lectin crosslinks and binds to sulfatides with a high affinity, in a process that could result in unwanted signaling events in the body, making this another soybean lectin antinutritional pathway. This discovery could also lead to the design of suitable additives to target the sulfatide binding site on soybean lectin and prevent soybean lectin from binding to sulfatide in the body. It is recommended that this newly discovered soybean lectin-sulfatide anti-nutritional pathway should also be considered when assessing the nutritional quality of soybean meal. Taken together, this research provides alternative solutions that would lead to the enhancement of the nutritional quality of soybean meal and the mitigation of the adverse effects of soybean's anti-nutrients."]},{"key":"dc:description.abstractgeneral","label":"General Abstract","values":["Soybean, which is a highly nutritious crop also contains some anti-nutrients like lectin and trypsin inhibitors that impede nutrient utilization in the body, leading to stunted growth and sometimes pancreatic hypertrophy. The presence of these anti-nutrients in soybean makes the processing of soybean before human or animal consumption a necessity. The conventional method of soybean processing which involves moist heating of soybean is relatively inefficient as it leads to the loss of nutrients, and processed soybean contains residual amount of anti-nutrients. This research work aimed to find alternative solutions to mitigate the adverse effects of soybean anti-nutrients by studying the way they interact and carry out their anti-nutritional functions in the body. Part of the alternative solutions to conventional soybean meal processing that this research work offers include the identification of soybean meal lines that are suitable for soybean crossbreeding to produce hybrid soybean meal lines that would require minimal processing before consumption, thereby, preventing the loss of nutrients that results from excessive soybean processing. We also revealed critical information that would lead to the design of additives that can be introduced into soybean meal to make soybean's anti-nutrients non-functional. Furthermore, we discovered a new way in which lectin interacts in the body to carry out its anti-nutritional effect. This provides a new parameter to consider during the quality control check on the nutritional quality of soybean meal before release for consumption."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Doctor of Philosophy"]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["ETD"]},{"key":"dc:title","label":"Title","values":["Molecular characterization of soybean meal trypsin inhibitors and lectins as a basis for developing approaches to mitigate their anti-nutritional effects"]}]}],"canonical_facts":{"dc:contributor.committeechair":["Capelluto, Daniel G."],"dc:contributor.committeemember":["Schubot, Florian David","Welborn, Valerie","Mevers, Emily Elizabeth"],"dc:contributor.department":["Chemistry"],"dc:creator":["Okedigba, Ayoyinka Oluwaseun"],"dc:date.accessioned":["2025-12-04T09:00:22Z"],"dc:date.available":["2025-12-04T09:00:22Z"],"dc:date.issued":["2025-12-03"],"dc:description.abstract":["Soybean is a crop that is widely eaten because of its nutritive properties. However, soybean cannot be consumed in its raw form because it contains anti-nutrients like trypsin inhibitors which bind to serine proteases (chymotrypsin, elastase and trypsin) and restrain their digestive functions leading to indigestion and stunted growth. It also contains lectin which binds to intestinal cells containing N-acetyl galactosamine (GalNAc) to impede nutrient uptake into the blood. Soybean needs to be processed to get rid of these anti-nutrients before human or animal consumption. The conventional method of soybean processing which involves the moist heat treatment of soybeans is time and energy demanding, leads to the loss of nutrients, and residual amount of anti-nutrients are retained in the processed soybean meal, making this process relatively inefficient. This research work aimed to provide alternative solutions to the conventional soybean processing methods. Using quick purification techniques, we isolated TIs [Bowman-Birk trypsin inhibitor (BBTI) and Kunitz trypsin inhibitor (KTI)] and lectin from soybean meal. Using biophysical techniques, we characterized the interactions between soybean anti-nutrients and their host ligands to offer alternative solutions to improve the nutritional quality of soybean meal. The known anti-nutritional pathway of soybean lectin involves its binding to GalNAc containing intestinal cells; however, we believe soybean lectin could be also targeting another ligand called sulfatide besides GalNAc in the small intestine due to some similarities between soybean lectin and galectin-4, a mammalian lectin that binds to sulfatide in the small intestine. Hence, we also explored soybean lectin to sulfatide interactions to find a new soybean lectin anti-nutritional pathway. Results from a soybean meal cultivar showed that KTI had a binding preference for chymotrypsin, while BBTI preferred binding to trypsin and elastase. This provides insights into the unique roles that both TIs play in soybean and why the plant retains both. After screening TIs from several soybean meal lines for their affinity to trypsin, we identified BBTI soybean lines with ~4 to 6-fold lower affinity for trypsin and a KTI soybean line with ~8-fold lower affinity for trypsin, compared to the standard BBTI and KTI, respectively. These promising soybean meal lines have been designed for soybean crossbreeding to produce hybrid soybean meal lines that would require little to no processing before animal consumption. We also identified the KTI amino acid sequence motif, SPLHALFI as a suitable motif for gene editing to produce soybean meal lines with improved nutritional qualities. After screening GalNAc and GalNAc analogs against lectin to find a higher lectin affinity ligand than GalNAc that could be introduced as an additive in soybean meal to bind to lectin and prevent lectin from binding to GalNAc containing intestinal cells, we found no higher affinity ligand than GalNAc for lectin. However, we found out that a suitable substitution on the anomeric carbon of GalNAc would lead to the design of higher lectin affinity ligands than GalNAc that could serve as additives in soybean meal to mitigate lectin adverse effects. We also found out that soybean lectin crosslinks and binds to sulfatides with a high affinity, in a process that could result in unwanted signaling events in the body, making this another soybean lectin antinutritional pathway. This discovery could also lead to the design of suitable additives to target the sulfatide binding site on soybean lectin and prevent soybean lectin from binding to sulfatide in the body. It is recommended that this newly discovered soybean lectin-sulfatide anti-nutritional pathway should also be considered when assessing the nutritional quality of soybean meal. Taken together, this research provides alternative solutions that would lead to the enhancement of the nutritional quality of soybean meal and the mitigation of the adverse effects of soybean's anti-nutrients."],"dc:description.abstractgeneral":["Soybean, which is a highly nutritious crop also contains some anti-nutrients like lectin and trypsin inhibitors that impede nutrient utilization in the body, leading to stunted growth and sometimes pancreatic hypertrophy. The presence of these anti-nutrients in soybean makes the processing of soybean before human or animal consumption a necessity. The conventional method of soybean processing which involves moist heating of soybean is relatively inefficient as it leads to the loss of nutrients, and processed soybean contains residual amount of anti-nutrients. This research work aimed to find alternative solutions to mitigate the adverse effects of soybean anti-nutrients by studying the way they interact and carry out their anti-nutritional functions in the body. Part of the alternative solutions to conventional soybean meal processing that this research work offers include the identification of soybean meal lines that are suitable for soybean crossbreeding to produce hybrid soybean meal lines that would require minimal processing before consumption, thereby, preventing the loss of nutrients that results from excessive soybean processing. We also revealed critical information that would lead to the design of additives that can be introduced into soybean meal to make soybean's anti-nutrients non-functional. Furthermore, we discovered a new way in which lectin interacts in the body to carry out its anti-nutritional effect. This provides a new parameter to consider during the quality control check on the nutritional quality of soybean meal before release for consumption."],"dc:description.degree":["Doctor of Philosophy"],"dc:format.medium":["ETD"],"dc:identifier.other":["vt_gsexam:44800"],"dc:identifier.uri":["https://hdl.handle.net/10919/139821"],"dc:language.iso":["en"],"dc:publisher":["Virginia Tech"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:subject":["Soybean meal","Bowman-Birk trypsin inhibitor","Kunitz trypsin inhibitor","serine proteases","lectin","N-acetyl galactosamine","sulfatides"],"dc:title":["Molecular characterization of soybean meal trypsin inhibitors and lectins as a basis for developing approaches to mitigate their anti-nutritional effects"],"dc:type":["Dissertation"],"thesis:degree_discipline":["Chemistry"],"thesis:degree_level":["doctoral"],"thesis:degree_name":["Doctor of Philosophy"],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:19:42Z"}